PA5-29760

antibody from Invitrogen Antibodies

Targeting: RPS6 S6

Antibody data

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Validation data

Product number: PA5-29760 - Provider product page
Provider: Invitrogen Antibodies
Product name: S6 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: HeLa, HepG2, Mouse brain. Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (96%), Drosophila (84%), Xenopus laevis (96%), Dog (100%), Chicken (98%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Mouse, Zebrafish
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 2.34 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RPS6 protein structure - PA5-29760 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of RPS6 using 30 µg of A) 293T and B) A431 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a RPS6 polyclonal antibody (Product # PA5-29760) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of RPS6 in Various whole cell extracts (30 µg). Samples were separated by 12% SDS-PAGE and the membrane was probed with RPS6 Polyclonal antibody (Product # PA5-29760) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using S6 Polyclonal Antibody (Product # PA5-29760). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 12% SDS PAGE. S6 Polyclonal Antibody (Product # PA5-29760) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using S6 Polyclonal Antibody (Product # PA5-29760). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with S6 Polyclonal Antibody (Product # PA5-29760) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using S6 Polyclonal Antibody (Product # PA5-29760). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with S6 Polyclonal Antibody (Product # PA5-29760) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of S6 was achieved by transfecting HeLa with S6 specific siRNAs (Silencer® select Product # S12270, S12272). Western blot analysis (Fig. a) was performed using whole cell extracts from the S6 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with S6 Polyclonal Antibody (Product # PA5-29760, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to S6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-S6 Polyclonal Antibody(Product # PA5-29760) and a 31kDa band corresponding to S6 was observed across all cell lines tested. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), Jurkat (Lane 4), NTERA2 cl.D1 (Lane 5) and U-87 MG (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of RPS6 in paraformaldehyde-fixed A431 cells using a RPS6 polyclonal antibody (Product # PA5-29760) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of S6 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: S6 Polyclonal Antibody (Product # PA5-29760) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of S6 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with S6 Polyclonal Antibody (Product # PA5-29760) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytosolic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.